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Sensitive and efficient monitoring of food-borne bacteria is of great importance for food safety control. Herein, a novel biosensor for highly sensitive detection of Staphylococcus aureus (S. aureus) was constructed by combining hybridization chain reaction (HCR) and nicking enzyme. Different from the upstream-downstream based circuit, the proposed biosensor integrated HCR circuit and three-way DNA junction nicking enzyme assisted signal amplification (3WJ-NEASA) into a virtuous circle of promotion. In the HCR-mediated 3WJ-NEASA sensing strategy, target DNA of S. aureus initiated the self-assembly between HCR hairpins (H1 and H2), which exposed the gap to capture molecular beacon (MB) and construct the 3WJ structure. Meanwhile, MB increased the stability of HCR nanowires and enhanced the efficiency of the HCR circuit, and thus more 3WJ-NEASA circuits were generated in HCR nanowires. Benefiting from the synergistic amplification coupling HCR and 3WJ-NEASA, this isothermal biosensor can detect as low as 6.7 pM of target DNA in one step within only 30 min. Furthermore, the HCR-mediated 3WJ-NEASA assay has been applied in the detection of S. aureus with a limit of detection (LOD) as low as 1.2 × 101 cfu mL−1, and has exhibited reliable practicability in spiked milk. It is the first time that a DNA biosensor combining HCR and 3WJ-NEASA for dual signal amplification was developed and has been adopted to the sensitive analysis of food-borne bacteria. Additionally, this strategy can serve as a universal platform for monitoring other analytes, and therefore possesses broad application prospects in food safety and environmental monitoring. [ABSTRACT FROM AUTHOR]

Polyaniline (PANI) is one of the most studied conducting polymers. It could adopt a number of chemical forms depending on the redox states of the assay solution. It was observed that the emeraldine base form of PANI could be reduced by common antioxidants (such as glutathione and ascorbic acid), which resulted in clear changes of the solution color and the UV–vis absorption spectra. In addition, it was observed that the fluorescence of CdTe quantum dots could be efficiently quenched by PANI, and the fluorescence quenching could not take place by the addition of antioxidants. The intensity of the quantum dots fluorescence could be directly related to the amount of antioxidant added to the assay solution, thus a sensitive fluorescent turn-on sensor for antioxidant was established. The detection limits for glutathione and ascorbic acid were estimated to be 50 nM and 100 nM, respectively. Our assay is simple, sensitive, and label-free, we envision it could facilitate the antioxidant sensing related applications. [ABSTRACT FROM AUTHOR]

Based on the controlled aggregation of quantum dots (QDs), a valid, reliable, and label-free fluorescence turn-on strategy is established for the detection of alkaline phosphatase activity. The aqueous solution of anionic QDs exhibits intense fluorescence. However, the addition of cationic polymer (poly-1) significantly quenched the fluorescence of the QDs via their induced aggregation. While short 3′-phosphorylated DNA (DNA-P) could not be extended by terminal deoxynucleotidyl transferase (TdT) and therefore, fluorescence of the QDs was recovered negligibly. The effective elimination of phosphate group of DNA-P in the presence of alkaline phosphatase (ALP) produced 3′-OH termini and the resulting DNA could be sufficiently extended by TdT. The presence of greater binding strength between the elongated DNA and poly-1 is very crucial to compete with the poly-1/QDs aggregates and release the QDs. Turned-on fluorescence emission is observed due to the efficient release of the QDs. A novel strategy for alkaline phosphatase detection is therefore established. Our method is quite sensitive and selective, as low as 0.1 mU/mL ALP can be easily detected. [ABSTRACT FROM AUTHOR]

SARS-CoV-2 isolation from cold-chain food products confirms the possibility of outbreaks through cold-chain food products. RNA extraction combined with RT-PCR is the primary method currently utilized for the detection of SARS-CoV-2. However, the requirement of hours of analytical time and the high price of RT-PCR hinder its worldwide implementation in food supervision. Here, we report a fluorescence biosensor for detection of SARS-CoV-2 N protein. The fluorescence biosensor was fabricated by aptamer-based conformational entropy-driven circuit where molecular beacon strands were labeled with graphitic carbon nitrides quantum dots@Zn-metal-organic framework (g-CNQDs@Zn-MOF) and Dabcyl. The detection of the N protein was achieved via swabbing followed by competitive assay using a fixed amount of N-48 aptamers in the analytical system. A fluorescence emission spectrum was employed for the detection. The detection limit of our fluorescence biosensor was 1.0 pg/mL for SARS-CoV-2 N protein, indicating very excellent sensitivity. The fluorescence biosensor did not exhibit significant cross-reactivity with other N proteins. Finally, the biosensor was successfully applied for the detection of SARS-CoV-2 N protein in actual cold-chain food products showing same excellent accuracy as RT-PCR method. Thus, our fluorescence biosensor is a promising analytical tool for rapid and sensitive detection of SARS-CoV-2 N protein. • An aptamer-based sensor could detect of SARS-CoV-2 in food products within 30 min. • Conformational entropy-driven circuit was designed for signal amplification. • g-CNQDs@Zn-MOF was utilized to improve stability and reproducibility of biosensor. • This strategy unlocks a route for the development of sensitive analytical tools. [ABSTRACT FROM AUTHOR]